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Bacillus sp. ATCC 29669 was isolated from microbial fallout in clean rooms during the assembly of the Viking Spacecraft missions to Mars, making it a potential contamination concern for outbound space missions. Spores from this bacterial strain were found to be thirty times more resistant to dry heat than B. atrophaeus. Spore inactivation rates under vacuum controlled humidity were faster than rates obtained under ambient humidity. Inactivation rates for these heat resistant spores are important considerations for planetary protection implementation where temperature, time and humidity conditions are used to estimate the effectiveness of dry heat microbial reduction (DHMR) procedures.

This study of samples collected from Mars 01 Orbiter was conducted to gain a better understanding and practical experience in methods to process and preserve samples intended for planetary protection analysis. Samples were evaluated for the viable growth of microbes, the molecular biomarker adenosine triphosphate (ATP), and the presence of lipopolysaccharide, a bacterial cell wall component. Losses were observed in the number of viable microbes after freezing as well as in the detectable lipopolysaccharide. Two independent studies of pooled cleanroom samples demonstrate good ATP recovery and consistent values after freezing at −20 °C.

In order to meet microbial reduction requirements for all Mars in-situ life detection and sample return missions, entire planetary spacecraft (including planetary entry probes and planetary landing capsules) may have to be exposed to a qualified sterilization process. At JPL, we are developing a low temperature (~45°C) vapor phase hydrogen peroxide sterilization process. This process is currently being used by the medical industry and its effectiveness is well established. In order to effectively and safely apply this technology to sterilize a spacecraft, which is made out of various man-made materials and electronic circuit boards, the following technical issues need to be resolved: 1. Efficacy of sterilization process. 2. Diffusion of H2O2 under sterilization process conditions into hard to reach places. 3. Materials and components compatibility with the sterilization process. 4. Development of methodology to protect (isolate) sensitive components (i.e. electronic ) from H2O2 vapor.